Glucose-regulated localization of Snf1–GFP and Snf4–GFP. Strains MCY4455 (wild type, WT) and MCY4458 (gal83Δ) were transformed with centromeric plasmids pOV84 and pOV76, expressing Snf1–GFP and Snf4–GFP, respectively. Cells were grown to mid-log phase on synthetic medium with 2% glucose (Glu) and shifted to 2% glycerol (Gly) for 20 min. (A) Protein extracts were prepared from the indicated cultures, separated by SDS-PAGE, and immunoblotted with anti-GFP antibody. (B,C) Wild-type (B) and gal83Δ mutant (C) cells expressing the indicated proteins were examined by fluorescence microscopy. (GFP) green fluorescent protein.